de Souza Filho J P, Correa Z M S, Marshall J C, Anteka E, Coutinho A B, Martins M C, Burnier M N
The Henry C Witelson Ocular Pathology Laboratory, McGill University, Montreal, Canada.
Eye (Lond). 2006 May;20(5):598-601. doi: 10.1038/sj.eye.6701938.
To examine the effect of nepafenac, a selective cyclooxygenase-2 (COX-2) inhibitor, on the proliferation rate of two human retinoblastoma (Rb)cell lines.
Two human Rb cell lines (WERI-RB and Y79) were cultured. COX-2 expression in these cell lines was verified by immunocytochemical analysis of cytospin sections and Western blotting. An MTT-based proliferation assay was used to compare Rb cell growth with and without amfenac, the active metabolite of nepafenac. The averaged results per condition were recorded. The Student's t-test was used to compare results from the cells cultured with and without amfenac.
The Y79 cell line showed a higher proliferative rate than the WERI-RB cell line. Both cell lines were negative for COX-2 expression by immunocytochemical analysis; however, both cell lines were positive for COX-2 expression by Western blot. When amfenac was added to both of the cell lines, a statistically significant reduction in proliferation was observed in both cell lines. The two Rb cell lines were positive for COX-2 only in the Western blot, indicating that they probably express low levels of COX-2, which was undetectable by immunocytochemical analysis.
The selective, anti-COX-2 molecule amfenac inhibited proliferation of both tested Rb cell lines. Further trials should be undertaken to study the effect of selective COX-2 inhibitors on Rb.
研究选择性环氧化酶-2(COX-2)抑制剂奈帕芬酸对两种人视网膜母细胞瘤(Rb)细胞系增殖率的影响。
培养两种人Rb细胞系(WERI-RB和Y79)。通过细胞涂片免疫细胞化学分析和蛋白质印迹法验证这些细胞系中COX-2的表达。使用基于MTT的增殖试验比较有和没有奈帕芬酸的活性代谢物氨苯芬酸情况下Rb细胞的生长情况。记录每种条件下的平均结果。采用Student t检验比较有和没有氨苯芬酸培养的细胞结果。
Y79细胞系的增殖率高于WERI-RB细胞系。通过免疫细胞化学分析,两种细胞系COX-2表达均为阴性;然而,通过蛋白质印迹法,两种细胞系COX-2表达均为阳性。当向两种细胞系中加入氨苯芬酸时,观察到两种细胞系的增殖均有统计学意义的降低。两种Rb细胞系仅在蛋白质印迹法中COX-2呈阳性,表明它们可能表达低水平的COX-2,免疫细胞化学分析无法检测到。
选择性抗COX-2分子氨苯芬酸抑制了两种受试Rb细胞系的增殖。应进行进一步试验以研究选择性COX-2抑制剂对Rb的影响。